Method for preparing the anhydrous crystalline form of isoniazid-derived hydrazone, thus produced crystalline polymorph of the anyhydrous form, use thereof for the treatment of alzheimer&#39;s disease, parkinsonism and other neurodegenerative disorders, and pharamceutical composition

ABSTRACT

A method for preparing the anhydrous crystalline form of isoniazid-derived hydrazone (8-hydroxyquinoline-2-carboxaldehyde isonicotinoyl hydrazone), to the thus produced polymorph and to the use thereof for the treatment of Alzheimer&#39;s disease and parkinsonism, inter alia, and to a pharmaceutical composition.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims priority on and the benefit of U.S. patent application Ser. No. 15/106,181 having a filing date of 17 Jun. 2016, which claims priority on and the benefit of and is the US National Phase under 35 USC 371 of International Application No. PCT/BR2014/000186 having an international filing date of 6 Jun. 2014, which claims priority on and the benefit of Brazilian Patent Application No. 10 2013 033006-0 having a filing date of 20 Dec. 2013.

BACKGROUND OF THE INVENTION Field of the Invention

The present invention regards the preparation method of the anhydrous crystalline form of an isoniazid-derived hydrazone (namely, 8-hydroxyquinoline-2-carboxaldehyde isonicotinoyl hydrazone, or INHHQ), the polymorph produced and its use, in any pharmaceutical composition, for the treatment of Alzheimer's and Parkinson's diseases, as well as other neurodegenerative disorders.

Prior Art

With the general increase in life expectancy recorded in recent times, especially in developed countries, the prevalence of certain neurodegenerative diseases has been observed. Alzheimer's disease (AD), first characterized in 1906 by the german neuropathologist Alois Alzheimer, is currently the most common one. It is a primarily age-related disease and the most common cause of dementia in older people. Dementia is characterized by progressive loss of memory and cognitive functions, among other diagnostic criteria, being most of them present in the Diagnostic and Statistical Manual of Mental Disorders and described on the National Institute of Neurological and Communicative Disorders Association.

The disease has two general classifications: 1) late onset, which occurs with the highest incidence at about 60 years of age, and 2) early onset, occurring around 40 years. In the US and Great Britain, it represents about 50% of cases of dementia, being estimated that it is the fourth leading cause of deaths of elderly in these countries. Regarding the neuropathological aspect, patients show diffuse cortical atrophy, presence of senile plaques and neurofibrillary tangles, neurovascular degeneration and neuronal loss.

These senile plaques are characterized mainly by the presence of fibrillar deposits of β-amyloid peptide (Aβ), consisting of approx. 40 amino-acid residues. It has been observed high concentration of physiological metal ions such as Zn²⁺ and Cu²⁺, in these plaques, which is considered indicative that the interaction of Aβ with these biometais is at the heart of events that lead to aggregation and toxicity of this peptide. The ferric ions, in turn, have also been related to the aggregation of neurofibrillary tangles, in addition to contributing to the oxidative processes that occur in the nerve cells of the body. Something similar occurs in Parkinson's disease with the proteinα-synuclein.

The already approved drugs for the treatment of AD are intended to combat deficits associated with reduced cerebral function and fall into two classes: acetylcholinesterase inhibitors and inhibitors of NMDA (N-Methyl-D-Aspartate) receptors. Such drugs seem to act in enhancing the remain of the cognitive function, however, are not able to prevent the progression of the disease, being, therefore, important the development of new therapeutic agents that hold the advance of neurodegeneration as well as, as far as possible, promote their regression. In this sense, an interesting approach is to obtain compounds that inhibit, specifically, abnormal metal-protein interactions. This class of drugs is known as MPACs (metal-protein attenuating compounds) and relate to the allocation and distribution normalization of physiological metal ions.

A classic example is the clioquinol (CQ or PBT1), a substance belonging to the group of 8-hydroxyquinolines, which, however, was abandoned due to certain unwanted side effects, such as subacute myelo-optic neuropathy.

Structure of clioquinol (5-chloro-7-iodo-8-hydroxyquinoline).

8-hydroxyquinoline-2-carboxaldehyde isonicotinoyl hydrazone (or INHHQ) was first described in a series of publications from the year 2009, in which its interactions with some rare earths ions, namely, dysprosium(III), europium(III), holmium(III), neodymium(III), and ytterbium(III) were examined. Erbium(III) and terbium(III) complexes were reported in 2010, while the samarium(III) compound was described in 2011. In those works, its rare earths complexes, and not INHHQ itself, were proposed as potential anticancer drugs, since they bind to DNA through an intercalation mechanism, besides they possess antioxidant properties, scavenging hydroxyl and superoxide radicals. Although the method of preparation described by the authors of those papers, Liu & Yang, is similar to the one employed by us, some experimental details are different. Mainly, the use of acid catalysis (drops of concentrated hydrochloric acid) allow us to reduce the reflux period to just an hour. On the other hand, and more important yet, the use of methanol PA ACS (99.8%) in the recrystallization step, instead of the methanol:water 80:20 used by the Chinese authors, lead to the preparation of the anhydrous crystalline form of the compound, which is completely original. Although this is not commented in the original articles, data suggest that the Chinese authors worked with an hydrated form of INHHQ, different from the one obtained by us, which can be clearly seen in the major differences between the melting points of the compounds, as well as between the vibrational (FTIR and Raman) spectra of both samples. Besides, application of INHHQ in the treatment of neurodegenerative diseases is completely new.

The anhydrous crystalline form described herein is characterized by its melting point, elemental analysis, crystal structure (single-crystal diffraction), powder diffraction pattern, and vibrational spectra.

BRIEF SUMMARY OF THE INVENTION

In the search for new MPACs, more effective and safer for patients, a hydrazone derived from the mycobactericidal agent isoniazid was produced, characterized in its crystalline form and successfully tested in modulating in vitro the interaction between β-amyloid peptide (or α-synuclein)and the physiological metals copper and zinc, which can have great impact on the treatment of Alzheimer's and Parkinson's diseases, as well as in other related diseases.

BRIEF DESCRIPTION OF THE DRAWINGS

The present invention will hereinafter be illustrated with reference to the attached figures, which represent:

FIG. 1: ORTEP representation of the compound INHHQ, showing the identification scheme of the constituent atoms;

FIG. 2: Perspective views of INHHQ showing: (a) the hydrogen-bonding network, (b) the π-π and the O1-H12

π stacking interactions and (c) the 3D crystalline packing along the crystallographic axis a;

FIG. 3: Simulated and experimental X-ray diffractograms for INHHQ;

FIG. 4: Most stable calculated structure in the gas-phase (Conf. 5). Level of theory: B3LYP/6-311+G (d,p);

FIG. 5: FTIR (above) and Raman (below) spectra of INHHQ;

FIG. 6: 2D ¹H×¹⁵N HSQC contour plots showing that the addition of INHHQ (4 eq.) to a mixture containing the β-amyloid peptide and Zn(II) (1 eq.) leads to a partial intensity recovery for all Aβ signals, which can be considered as an indicative that the hydrazone INHHQ weakens Zn(II)-Aβ interactions;

FIG. 7: ¹H NMR spectra showing that the addition of INHHQ (4 eq.) to a mixture containingα-synuclein and Cu(II)/Cu(I) (1 eq.) leads to the recovery of the original methionine signals' position; it is worth noting that methionine residues are the interaction point of copper with α-synuclein, which constitutes an indicative that INHHQ weakens Cu(II) or Cu(I)-α-synuclein interactions.

DETAILED DESCRIPTION OF THE INVENTION

The isonicotinoyl hydrazone of 8-hydroxyquinoline-2-carboxaldehyde (or INHHQ) was synthesized, completely characterized in its anhydrous crystalline form and successfully tested in modulating the in vitro interaction between β-amyloid peptide (or α-synuclein) and the physiological metals copper and zinc, which can have applications in the treatment of Alzheimer's and Parkinson's diseases, amongst other neurodegenerative diseases (such as Huntington's).

Structure of INHHQ (8-hydroxyquinoline-2-carboxaldehyde isonicotionoyl hydrazone)

Connected to the 8-hydroxyquinoline portion, characteristic of CQ, INHHQ also contains the mycobactericidal drug isoniazid (INH), resulting in a potentially interesting hydrazone capable of coordinating metal ions of biological importance through its various N/O-donor sites. Furthermore, linking two molecules that have, individually, specific activity, creating a single species, has been an attractive approach to rational drug development, since a combination of the two original activities can be expected of the hybrid molecule. Besides, isoniazid-derived hydrazones are well known as iron chelating agents. However, to the best of our knowledge, there are no studies in the literature involving the coordination of INHHQ to any transition metal.

In the context of the treatment of AD and Parkinson's disease, the coordination capacity of the ligand to essential transition metals (such as copper, zinc and iron) must be better understood. As a first approach to this problem, we present the results of a spectroscopic vibrational (FTIR/Raman) and a single-crystal X-ray diffraction structural study on the 8-hydroxyquinoline-2-carboxaldehyde isonicotinoyl hydrazone, obtained through the process herein described and claimed, including its crystal structure, vibrational spectra and their full assignment by computational methods based on the Density Functional Theory (DFT).

Preparation Methodology of the Anhydrous Crystalline Form of 8-hydroxyquinoline-2-carboxaldehyde isonicotinoyl hydrazone

To 25 mL of a 8-hydroxyquinoline-2-carboxaldehyde (1.04 g, 6.0 mmol) solution in ethanol, 25 mL of ethanolic solution of isoniazid (0.82 g, 6.0 mmol) were dropwise added. Two drops of concentrated hydrochloric acid were added as a catalyst for the reaction. After refluxing for 1 h, the mixture was cooled to room temperature and the yellow precipitate obtained was filtered and vacuum dried. Next, the product was recrystallized in hot methanol methanol PA ACS (99.8%). A pale yellow crystalline solid was obtained after cooling and dried at room temperature. After few days, single-crystals of INHHQ were isolated from the mother liquor. Total yield of the process: 0.98 g (56%).

Characterization: m.p.: 246-249° C.

Elemental analysis: calculated for C₁₆H₁₂O₂N₄: C, 65.7%; H, 4.1%; N, 19.2%—found: C, 66.3%; H, 4.1%; N, 19.4%.

Main IR bands (KBr, cm⁻¹): 3396 (v OH); 3183 (v NH); 1656 (v C═O+β NH); 1647 (v C═N azomethine group); 1556 (v C═N e v C═C from the quinolinic ring) and 1545 (v C═C e v C═N from the pyridinic ring).

X-ray Diffraction Analysis

The X-ray diffraction (XRD) was performed using an appropriate single-crystal of INHHQ. The sample was measured on a diffractometer Enraf-Nonius Kappa-CCD with Mo K_(α) (λ=0.71073 Å) radiation. The unit cell parameters were based on all reflections. Data were collected at room temperature (293 K) using the computer program Collect being the integration and scaling of reflections made with Denzo—Scalepach system of HKL programs. The crystal structure was solved by direct method with SHELXS-97 and the atoms, except hydrogens, were anisotropically refined by the least squares method on F2 using the SHELXL-97 program. All aromatic and hydroxyl group's hydrogen atoms were placed in the calculated positions (C—H: 0.98 Å, O—H: 0.82 Å). Shift factors were taken as U(H)_(isot)=1.2/1.5 Uhost. The H atoms attached to the C7 carbon and N2 nitrogen were located in the Fourier difference map and refined freely. The computer programs ORTEP-3 and Mercury (version 2.3) were used to draw the structures. On the other hand, powder X-ray diffraction experiments were performed in a Bruker D8 Discover XRD equipment using copper radiation. Experimental conditions: tension of 40 kV and current of 40 mA. The Bragg-Brentano geometry was used.

Spectroscopic Analyses

IR spectra were obtained on a Perkin-Elmer 2000 FT-IR spectrometer, using KBr sampling. On the other hand, the Raman spectra of the solid sample were measured on a Perkin-Elmer 400 Station equipment, using the 785 nm line for excitation.

DFT Calculations

The first step was to conduct a search of the conformational space of 8-hydroxyquinoline-2-carboxaldehyde isonicotinoyl hydrazone using the semi-empirical PM3 method, as implemented in the SPARTAN'02 software, with the following set of parameters: Max-Confs=100; window=10 kcal mol⁻¹; model=PM3. A total of 12 different conformations were found. The lowest energy conformation was of 41.387 kcal mol⁻¹. From this distribution, 5 conformations were selected for analysis by DFT. These conformations will be called, from now on, from Conf. 1 to Conf. 5.

After the selection of these five conformations, a complete geometry optimization, without limitation, using the default convergence criteria and calculation of harmonic vibrational frequencies, was performed in gas phase, for each, using the Gaussian program package 03 Gauss. The three parameters of the exchange-correlation potential proposed by Becke with local and non-local correlations provided by Lee, Yang, and Parr (B3LYP functional) were selected with the basis set of triple zeta valence 6-311 +G (d,p). Thermal contributions to the Gibbs free energy and other state functions were calculated at 298.15 K and 1 atm. The vibrational frequencies were scaled by a factor of 0.9381 for a better comparison with the experimental data.

Molecular Structure Crystallographic Analysis

The main crystal parameters used, as well as data of information collected and structural refinement parameters are summarized in Table 1. INHHQ crystallizes in the orthorhombic system, space group Pbca. The asymmetric unit of INHHQ is shown in FIG. 1. The compound adopts an (E) configuration with respect to the C7=N3 bond of the hydrazonic group. The INHHQ molecule is almost planarin solid state (rms deviation=0.2701° for all atoms, except H) and shows an intramolecular H bond involving the phenolic hydroxyl and the quinolinic nitrogen group: the O1-H donor interacts with N4 [O1

N4=2,689 Å] receptor. In this process, a pseudo five-membered ring is formed. The distances and bond angles (Table 2) are not significantly different from those observed in similar compounds. The crystal packing is maintained by intermolecular H bonds involving the carbonyl oxygen O2 (acceptor) of a molecule and the N2-H group of the following molecule [moderate N2

O2^(i)=2,966 Å, symmetry codes: (i) −x+½, y−½, z], connecting the molecules of INHHQ in zigzag chains running parallel to the crystallographic b axis (FIG. 2(a)). The molecules in each chain are interconnected by cross-stacking π-π interactions involving the quinoline rings. The calculated centroid-centroid distance is equal to 3.8303(9) Å. The adjacent chains are interconnected by O1-H12

π interactions (FIG. 2(b)), being the distance H12-centroid (N4-C8-C9-C10-C11-C12) of 3.5339(17) Å [symmetry code: −½+x, y, ½−z]. As a result of this last interaction, zigzag columns run parallel to the crystallographic axis a (FIG. 2(c)).

TABLE 1 Selected crystallographic data for INHHQ Empirical formula C₁₆H₁₂O₂N₄ Molecular weight 292.30 Collecting temperature 293(2) K. Wave length 0.71073 Å Crystalline system Orthorhombic Space group Pbca Unit cell dimensions a = 17.0761(4) Å  b = 8.25480(10) Å c = 19.3549(4) Å  Cell volume 2728.26(9) Å³ Z 8 ρ(calculated) 1.423 g cm⁻³ Absorption coefficient 0.098 mm⁻¹ F(000) 1216 Crystal size utilized 0.484 × 0.236 × 0.171 mm³ θ range used in collect 2.94 a 27.48° Index ranges −22.19; −10.10; −25.23 Collected reflections 27142 Unique 3100 [R_((int)) = 0.1167] Completeness to θ = 27, 48° 99.0% Absorption correction None Refinement method “Full-matrix least-squares on F^(2”) Computational programs ^(a) COLLECT, HKL Denzo and Scalepack, SHELXS−97, SHELXL−97 Data/restraints/parameters 3100/0/208 Goodness-of-fit on F² 1.042 Final Rindexes [I > 2σ (I)] R₁ = 0.0517, wR₂ = 0.1188 R index (all data) R₁ = 0.0968, wR₂ = 0.1427 Largest diff. peak and hole 0.198 and −0.234 e Å⁻³ ^(a) Used for data collection, data processing, structure solution, and structure refinement, respectively.

TABLE 2 Main distances and bond angles, experimental and theoretical (Conf. 5), for INHHQ. The crystallographic identification scheme of the atoms was followed Experi- Experi- mental Calculated mental Calculated Distances (Å) Distances (Å) O2—C6 1.2268(18) 1.211 N4—C12 1.362(2) 1.355 N2—C6 1.345(2)  1.388 N3—C7 1.270(2) 1.280 N2—N3 1.3862(18) 1.353 O1—C13 1.358(2) 1.351 N4—C8 1.319(2)  1.325 N1—C1 1.333(2) 1.335 Angles (°) Angles (°) C6—N2—N3 118.44(13) 120.9 N2—C6—C5 115.04(13) 114.0 C8—N4—C12 117.83(13) 118.6 C1—N1—C2 116.20(15) 117.2 C7—N3—N2 115.54(13) 117.4 O1—C13—C12 118.65(15) 118.8 Dihedral angles (°) Dihedral angles (°) C8—C7—N3—N2 −177.63 179.7 N3—N2—C6—C5 −178.62 −178.6 C7—N3—N2—C6 −163.79 −175.4 N2—C6—C5—C3 144.93 −151.6

Moreover, an analysis of the INHHQ diffraction peaks was performed using Bragg-Brentano geometry (Table 3). The ultimate goal is to characterize, unambiguously, the obtained polymorph. The crystal structure-based simulated diffractogram in the range of 5<2θ<70°, was also calculated. FIG. 3 displays the superposition of the experimental (red) and simulated (blue) diffractograms.

TABLE 3 Information on the diffraction peaks of INHHQ for 2θranging from 10.5 to 70.5° Interplanar Structure Plane (h, k, l) Multiplicity distance (Å) factor h k l m d 2 theta (°) F² 1 0 0 2 8.34877 10.58783 1128.338 0 1 0 2 7.07920 12.49361 331.562 0 1 1 4 5.94931 14.87873 4309.398 1 0 −2 2 5.93863 14.90564 3564.964 1 −1 −1 4 5.50433 16.08920 1058.563 0 0 2 2 5.48852 16.13588 185.920 1 1 0 4 5.39942 16.40394 3734.208 1 −1 −2 4 4.54974 19.49495 3562.456 1 1 1 4 4.37713 20.27167 81.922 2 0 −2 2 4.37659 20.27421 2544.993 0 1 2 4 4.33757 20.45853 131.263 2 0 0 2 4.17439 21.26742 743.827 2 −1 −1 4 3.87923 22.90666 5227.304 1 0 2 2 3.87110 22.95541 2615.373 2 −1 −2 4 3.72262 23.88427 1558.836 2 1 0 4 3.59579 24.73979 177.345 0 2 0 2 3.53960 25.13890 1609.181 1 −1 −3 4 3.53090 25.20189 4821.653 1 1 2 4 3.39646 26.21681 4682.200 0 2 1 4 3.36879 26.43604 783.016 1 −2 −1 4 3.28146 27.15290 1838.082 1 2 0 4 3.25881 27.34522 1594.341 2 −1 −3 4 3.25355 27.39035 1187.558 0 1 3 4 3.25049 27.41659 1027.571 3 0 −2 2 3.08971 28.87348 81.686 2 1 1 4 3.08859 28.88417 2178.500 1 −2 −2 4 3.04049 29.35123 126085.008 1 0 −4 2 3.03231 29.43221 46210.547 1 2 1 4 2.98732 29.88574 3959.848 0 2 2 4 2.97465 30.01599 23814.010 2 0 −4 2 2.96931 30.07124 20257.656 3 −1 −2 4 2.83175 31.56923 1316.031 2 −2 −1 4 2.81367 31.77739 2147.033 3 −1 −1 4 2.78865 32.07025 962.328 1 −1 −4 4 2.78737 32.08534 5017.017 2 0 2 2 2.78466 32.11737 5954.872 3 0 0 2 2.78292 32.13796 198029.766 2 −2 −2 4 2.75217 32.50702 166846.703 0 0 4 2 2.74426 32.60332 78569.977 2 −1 −4 4 2.73820 32.67747 7683.784 2 2 0 4 2.69971 33.15667 5524.309 3 −1 −3 4 2.69759 33.18355 5725.885 1 1 3 4 2.69411 33.22766 3144.983 1 −2 −3 4 2.67192 33.51166 5080.103 1 2 2 4 2.61222 34.30092 184684.922 3 0 −4 2 2.61018 34.32857 75081.414 2 1 2 4 2.59139 34.58541 10269.332 3 1 0 4 2.58999 34.60470 3123.402 0 1 4 4 2.55873 35.04100 3632.206 2 −2 −3 4 2.54551 35.22898 128.840 0 2 3 4 2.54404 35.24991 2518.650 2 2 1 4 2.46414 36.43237 9364.604 3 −1 −4 4 2.44902 36.66531 6171.995 0 3 0 2 2.35973 38.10499 2742.504 3 −2 −2 4 2.32767 38.65065 49774.492 1 0 4 2 2.32176 38.75283 27728.582 4 0 −2 2 2.31874 38.80530 8126.180 3 1 1 4 2.31772 38.82317 7422.130 0 3 1 4 2.30703 39.01032 1802.321 3 −2 −1 4 2.30355 39.07167 4033.272 1 −2 −4 4 2.30283 39.08438 4588.360 2 −1 −5 4 2.30097 39.11728 4454.373 1 −3 −1 4 2.27839 39.52090 11520.123 2 −2 −4 4 2.27487 39.58469 7746.600 1 −1 −5 4 2.27151 39.64563 598.446 1 3 0 4 2.27077 39.65907 2802.268 3 −2 −3 4 2.25142 40.01449 3656.189 1 2 3 4 2.24939 40.05204 2882.448 1 1 4 4 2.20614 40.87197 4367.143 4 −1 −2 4 2.20355 40.92216 1606.162 1 −3 −2 4 2.19295 41.12888 5896.017 2 2 2 4 2.18856 41.21509 90978.344 4 0 −4 2 2.18829 41.22042 43250.957 3 2 0 4 2.18772 41.23171 13279.063 4 −1 −3 4 2.18705 41.24487 37423.781 2 1 3 4 2.18427 41.29979 20309.209 1 3 1 4 2.17275 41.52890 27249.758 0 2 4 4 2.16879 41.60831 20608.090 3 −1 −5 4 2.16859 41.61228 15481.041 0 3 2 4 2.16786 41.62688 3233.373 4 −1 −1 4 2.13480 42.30230 2642.791 3 0 2 2 2.13258 42.34844 20.261 2 −3 −1 4 2.10312 42.97079 2666.499 3 −2 −4 4 2.10076 43.02150 1828.689 0 1 5 4 2.09689 43.10495 17605.400 4 −1 −4 4 2.09069 43.23919 2405.904 4 0 0 2 2.08719 43.31522 1974.789 2 −3 −2 4 2.07706 43.53728 2901.627 2 3 0 4 2.05423 44.04625 3037.751 3 1 2 4 2.04194 44.32549 1874.683 1 −3 −3 4 2.04191 44.32613 1474.088 2 0 −6 2 2.03690 44.44102 2853.471 3 2 1 4 2.01612 44.92393 1408.763 1 3 2 4 2.01489 44.95273 793.879 2 −2 −5 4 2.00506 45.18528 8021.569 4 1 0 4 2.00199 45.25840 3930.259 1 −2 −5 4 1.98548 45.65592 1014.055 2 −3 −3 4 1.98380 45.69690 16302.775 0 3 3 4 1.98310 45.71378 11086.845 3 0 −6 2 1.97954 45.80066 5989.497 1 0 −6 2 1.97817 45.83434 9677.354 2 −1 −6 4 1.95748 46.34679 2033.378 2 3 1 4 1.94455 46.67316 3814.112 4 −1 −5 4 1.94222 46.73259 2465.540 1 2 4 4 1.94138 46.75385 1044.347 4 −2 −2 4 1.93962 46.79894 71647.414 2 0 4 2 1.93555 46.90307 22593.117 4 −2 −3 4 1.92834 47.08917 2359.305 2 2 3 4 1.92643 47.13862 3496.081 3 −2 −5 4 1.91564 47.42022 3565.626 3 −1 −6 4 1.90641 47.66401 3852.061 1 −1 −6 4 1.90518 47.69668 126.165 4 −2 −1 4 1.89223 48.04363 2471.013 3 −3 −2 4 1.87535 48.50388 728.923 2 1 4 4 1.86703 48.73407 2582.125 0 2 5 4 1.86568 48.77150 687.112 3 −3 −1 4 1.86266 48.85561 371.683 1 −3 −4 4 1.86228 48.86626 3642.781 4 −2 −4 4 1.86131 48.89351 961.915 1 1 5 4 1.85776 48.99309 2897.553 2 −3 −4 4 1.84740 49.28595 2366.935 5 0 −2 2 1.83827 49.54725 191.546 4 1 1 4 1.83748 49.56990 893.848 3 −3 −3 4 1.83478 49.64793 34517.039 1 3 3 4 1.83368 49.67962 3586.474 4 0 −6 2 1.83168 49.73747 1341.276 0 0 6 2 1.82951 49.80075 5427.721 5 0 −4 2 1.82704 49.87241 2611.138 3 2 2 4 1.82666 49.88361 15035.101 2 3 2 4 1.80028 50.66584 852.734 3 3 0 4 1.79981 50.68000 7635.262 4 2 0 4 1.79790 50.73774 31.252 5 −1 −3 4 1.79778 50.74113 3492.466 3 1 3 4 1.79572 50.80350 1004.864 0 3 4 4 1.78922 51.00132 203.157 5 −1 −2 4 1.77926 51.30747 668.832 4 −1 −6 4 1.77329 51.49295 1637.459 0 1 6 4 1.77131 51.55465 738.791 0 4 0 2 1.76980 51.60187 118322.898 5 −1 −4 4 1.76908 51.62454 2002.556 2 −2 −6 4 1.76545 51.73845 267467.688 4 −2 −5 4 1.75423 52.09423 1767.949 3 −3 −4 4 1.75045 52.21513 447.095 0 4 1 4 1.74724 52.31836 2620.549 1 −4 −1 4 1.73470 52.72556 2457.296 1 4 0 4 1.73133 52.83614 1390.317 3 −2 −6 4 1.72771 52.95534 624.514 1 −2 −6 4 1.72680 52.98560 2870.138 5 −1 −1 4 1.71763 53.29042 3992.833 4 0 2 2 1.71625 53.33669 171.427 3 3 1 4 1.70057 53.86813 4463.794 5 −1 −5 4 1.69944 53.90672 1663.483 2 2 4 4 1.69823 53.94829 3506.858 1 −4 −2 4 1.69608 54.02211 2307.989 2 −3 −5 4 1.69392 54.09682 2349.578 1 2 5 4 1.69125 54.18921 1409.942 2 −1 −7 4 1.69108 54.19489 787.935 1 4 1 4 1.68669 54.34788 882.257 0 4 2 4 1.68440 54.42786 182.510 1 −3 −5 4 1.68206 54.50961 4158.873 3 −1 −7 4 1.68013 54.57753 2978.968 4 2 1 4 1.67591 54.72649 1026.918 5 0 0 2 1.66975 54.94516 2190.528 4 1 2 4 1.66794 55.01012 1131.238 1 3 4 4 1.65500 55.47716 297.480 4 −3 −2 4 1.65390 55.51703 92.102 2 −4 −1 4 1.65348 55.53225 2628.066 4 −3 −3 4 1.64689 55.77391 1703.457 2 3 3 4 1.64570 55.81771 14933.787 5 −2 −3 4 1.64562 55.82063 904.931 5 0 −6 2 1.64485 55.84915 11045.036 3 2 3 4 1.64404 55.87900 537.543 1 0 6 2 1.64248 55.93667 4104.017 2 −4 −2 4 1.64073 56.00177 6555.738 3 −3 −5 4 1.63896 56.06742 3117.231 1 −1 −7 4 1.63562 56.19224 7479.478 5 −2 −2 4 1.63138 56.35117 139974.406 2 4 0 4 1.62941 56.42557 2336.839 3 0 4 2 1.62858 56.45687 79060.617 4 −2 −6 4 1.62677 56.52512 231.031 0 2 6 4 1.62525 56.58301 200.360 5 1 0 4 1.62516 56.58630 1372.377 4 −3 −1 4 1.62423 56.62165 941.537 5 −2 −4 4 1.62352 56.64860 977.446 1 −4 −3 4 1.62324 56.65939 3828.351 2 1 5 4 1.62060 56.76011 661.635 1 4 2 4 1.60956 57.18482 2848.325 0 3 5 4 1.60734 57.27112 4687.052 4 −1 −7 4 1.60639 57.30817 785.019 4 −3 −4 4 1.60455 57.38024 979.594 5 −1 −6 4 1.60217 57.47322 229.132 1 1 6 4 1.59998 57.55912 756.968 2 −4 −3 4 1.59358 57.81228 6432.516 0 4 3 4 1.59322 57.82655 993.033 3 1 4 4 1.58712 58.06987 1480.846 5 −2 −1 4 1.58349 58.21592 1232.501 3 3 2 4 1.58219 58.26825 878.980 2 4 1 4 1.57302 58.64091 2737.882 5 −2 −5 4 1.56920 58.79766 1757.640 4 3 0 4 1.56339 59.03785 1224.589 2 −2 −7 4 1.56261 59.07000 858.527 3 −2 −7 4 1.55396 59.43196 388.934 6 0 −4 2 1.54485 59.81773 40226.578 4 2 2 4 1.54429 59.84161 132527.313 2 −3 −6 4 1.54191 59.94356 919.952 3 −4 −2 4 1.53571 60.21080 3337.484 4 −3 −5 4 1.53442 60.26657 2142.301 0 1 7 4 1.53103 60.41363 4066.389 3 −4 −1 4 1.52872 60.51473 530.741 1 −4 −4 4 1.52851 60.52394 23281.965 3 0 −8 2 1.52498 60.67867 13248.033 2 −4 −4 4 1.52025 60.88755 1880.706 1 −2 −7 4 1.51854 60.96338 1753.461 6 0 −2 2 1.51698 61.03271 2012.546 3 −3 −6 4 1.51658 61.05053 1466.016 5 1 1 4 1.51638 61.05931 161.170 2 0 −8 2 1.51616 61.06934 3710.299 1 −3 −6 4 1.51596 61.07813 2665.123 3 −4 −3 4 1.51319 61.20197 419.151 1 4 3 4 1.51257 61.22953 10115.472 6 −1 −3 4 1.51024 61.33418 363.616 5 2 0 4 1.51016 61.33806 617.463 6 −1 −4 4 1.50933 61.37515 989.558 4 1 3 4 1.50872 61.40297 297.840 2 2 5 4 1.50649 61.50344 588.937 2 3 4 4 1.49652 61.95818 590.113 4 −2 −7 4 1.49506 62.02536 423.413 2 4 2 4 1.49366 62.09007 106.506 3 4 0 4 1.49339 62.10246 123565.250 1 3 5 4 1.49174 62.17898 478.546 5 −2 −6 4 1.49166 62.18267 105324.078 5 −1 −7 4 1.49157 62.18692 1581.715 3 −1 −8 4 1.49078 62.22328 974.226 1 2 6 4 1.48989 62.26457 134633.219 0 4 4 4 1.48733 62.38401 83526.117 4 0 −8 2 1.48466 62.50882 50996.125 6 −1 −2 4 1.48331 62.57221 539.855 2 −1 −8 4 1.48254 62.60831 1032.783 4 3 1 4 1.48118 62.67218 6016.078 6 −1 −5 4 1.48072 62.69367 7655.220 3 2 4 4 1.47949 62.75186 5174.257 3 −4 −4 4 1.46483 63.45251 92852.609 1 0 −8 2 1.46062 63.65703 34382.762 5 −3 −3 4 1.46015 63.67986 19231.883 3 3 3 4 1.45904 63.73375 2083.547 6 0 −6 2 1.45886 63.74255 104.403 2 0 6 2 1.45666 63.85023 5589.541 4 −1 −8 4 1.45305 64.02798 501.111 5 −3 −2 4 1.45017 64.17003 1181.973 4 −3 −6 4 1.44693 64.33096 2199.269 0 3 6 4 1.44586 64.38455 3392.815 5 −3 −4 4 1.44464 64.44527 603.990 3 4 1 4 1.43522 64.91979 856.294 0 2 7 4 1.43374 64.99512 3941.112 6 −1 −1 4 1.43271 65.04772 9958.013 5 0 2 2 1.43178 65.09530 464.878 2 −4 −5 4 1.43122 65.12385 1343.613 1 −1 −8 4 1.43049 65.16140 2849.458 6 −1 −6 4 1.42884 65.24586 1632.203 2 1 6 4 1.42677 65.35220 113.830 1 −4 −5 4 1.42405 65.49281 2715.529 5 2 1 4 1.42169 65.61510 2686.276 6 −2 −3 4 1.41663 65.87928 845.347 5 −3 −1 4 1.41622 65.90063 1809.560 6 −2 −4 4 1.41588 65.91867 654.705 0 5 0 2 1.41584 65.92052 1197.549 4 2 3 4 1.41537 65.94542 244.526 3 1 5 4 1.41356 66.04055 4426.744 1 4 4 4 1.40751 66.36081 21200.830 4 −4 −2 4 1.40684 66.39668 396.245 5 −3 −5 4 1.40597 66.44291 29.975 5 0 −8 2 1.40574 66.45520 3142.987 0 5 1 4 1.40421 66.53712 7893.484 5 1 2 4 1.40336 66.58237 970.894 1 1 7 4 1.40288 66.60856 170.244 4 −4 −3 4 1.40251 66.62790 1310.150 2 4 3 4 1.40178 66.66737 2531.638 2 −3 −7 4 1.40123 66.69715 8505.064 5 −2 −7 4 1.40118 66.69978 401.337 3 −2 −8 4 1.40053 66.73479 5147.162 1 −5 −1 4 1.39767 66.88894 1543.957 3 −4 −5 4 1.39761 66.89248 1963.978 1 5 0 4 1.39591 66.98464 1123.055 3 −3 −7 4 1.39498 67.03543 1584.166 6 −2 −2 4 1.39432 67.07096 6423.831 2 −2 −8 4 1.39368 67.10576 2221.480 4 0 4 2 1.39233 67.17963 9756.798 6 −2 −5 4 1.39218 67.18803 123.361 6 0 0 2 1.39146 67.22715 61458.719 4 −4 −1 4 1.38844 67.39312 1208.544 4 3 2 4 1.38797 67.41877 146.200 5 −1 −8 4 1.37882 67.92709 1203.279 0 4 5 4 1.37785 67.98154 710.651 1 −5 −2 4 1.37724 68.01559 3062.095 4 −4 −4 4 1.37608 68.08057 27359.787 1 5 1 4 1.37219 68.30019 694.557 0 0 8 2 1.37213 68.30382 15200.482 0 5 2 4 1.37096 68.37016 828.054 1 −3 −7 4 1.36918 68.47118 3214.390 4 −2 −8 4 1.36910 68.47589 45.553 4 1 4 4 1.36616 68.64389 519.685 6 1 0 4 1.36534 68.69093 1607.419 5 3 0 4 1.36303 68.82355 2112.606 3 4 2 4 1.36190 68.88850 1753.492 6 −1 −7 4 1.36065 68.96085 2649.621 2 3 5 4 1.36033 68.97921 2510.422 2 −5 −1 4 1.35414 69.33989 3657.629 6 −2 −1 4 1.35205 69.46230 243.801 4 −3 −7 4 1.35190 69.47100 6915.945 1 −2 −8 4 1.35018 69.57237 1099.760 4 4 0 4 1.34986 69.59140 705.399 5 −3 −6 4 1.34938 69.61938 756.755 6 −2 −6 4 1.34879 69.65417 2227.248 1 3 6 4 1.34807 69.69667 318.911 2 −5 −2 4 1.34710 69.75419 61.678 0 1 8 4 1.34706 69.75684 1471.690 2 2 6 4 1.34705 69.75717 576.139 2 5 0 4 1.34082 70.12915 2349.962 3 3 4 4 1.34035 70.15689 35.647 1 −5 −3 4 1.33737 70.33644 2229.274 2 −4 −6 4 1.33596 70.42175 3337.051 3 2 5 4 1.33592 70.42439 156.499

Gas-Phase DFT Calculations

The INHHQ structure was optimized in gas phase, using the DFT methodology, level of theory B3LYP/6-311+G (d,p).

As described above, a total of 5 conformations with the smallest PM3 energy differences were selected to perform a DFT optimization and frequency calculations. Cis-trans isomerism was contemplated in this treatment. Energy values found for each of these structures indicated that there are, in fact, only 3 different conformations. Conf.5 is the one with the lowest free energy (ΔG) when compared to the other conformations. However, the energy differences between the structures are not appreciable, especially between the Conf. 1/2 with respect to the Conf. 5 (only 0.06 kcal mol⁻¹), indicating that, indeed, all five conformations are possible, particularly Conf. 5. This last one was chosen for further deeper studies.

It is shown (Table 2) that there is excellent agreement between the structural parameters found in the calculations (Conf. 5) and the refined X-ray structure and that, in turn, there is no significant difference between FIGS. 1 and 4. Crystallography shows that the phenolic hydrogen points in the direction of the quinolinic nitrogen, since a connection involving these H atoms is formed, producing a 2.691 Å donor-acceptor distance, in perfect agreement with the X-ray data (O1

N4=2.689 Å).

Vibrational Analyses

The experimental FTIR and Raman spectra of INHHQ, in the solid state, are shown in FIG. 5. The observed and calculated frequencies, as well as an attempt to assign the main bands, are given in Table 4.

TABLE 4 Assignment of the FTIR and Raman spectra of INHHQ (scale factor: 0.9381) Theoretical (cm⁻¹) B3LYP/6-311 + G(d, p) Experimental Un- (cm⁻¹) scaled Scaled IR Vibrational FTIRRaman (cm⁻¹) (cm⁻¹) Intensity Assignment 3396 m 3400 br 3662 3435 104.3584 v OH 3208 sh 3205 w — 3183 w — 3502 3285 5.5099 v NH 3148 sh — 3211/ 3012/ 1.8906/ v CH(Quin + Py)ip 3205 3007 3.2991 3073 sh 3081 w 3198 3000 9.2441 v CH(Quin)ip 3059 sh 3055 w 3185/ 2988/ 18.7464/ v CH(Quin + Py)op 3178 2981 9.9528 3042 w — — 3028 sh 3030 w — 3016 w — — — 2996 w — 2959 sh — — 2923 w — — 2852 w — 3048 2859 40.4835 vCH(Azomethine) 2835 sh — — 1656 s 1660 sh 1767 1658 350.6350 vC═O + βNH 1647 vs 1646 w 1673 1569 20.7477 v C═N (Azomethine) + v C═C(Quin) + βC—OH 1604 w 1603 vs 1659 1556 2.3397 v C═N (Azomethine) + v C═C(Quin) + βC—OH 1595 w 1595 s 1634 1533 5.5690 v C═C(Quin) 1556 m 1555 w 1597 1498 18.4149 v C═N(Quin) + v C═C(Quin) 1545 m — 1630 1529 14.2312 v C═C(Py) + vC═N(Py) 1507 m 1507 w 1555 1459 371.3416 βNH + βCH(Quin) 1490 w 1488 vw — 1465 m 1468 w 1541 1446 168.8474 Ring stretch(Quin) + βC—OH + β NH 1437 m 1435 s 1519 1425 6.8860 βCH(Py) 1407 w — — 1394 sh 1396 m — 1371 w 1371 m 1495 1402 176.2051 β C—OH + β NH + β CH(Quin) 1330 m 1329 m — 1299 s 1304 m 1360 1276 34.4059 β CH(Quin + Azomethine) + v C═N(Quin) + v C—OH 1280 m 1279 sh 1347 1264 31.7632 β NH + β CH(Azomethine + Py) + β C—OH 1270 sh — — 1252 m 1252 s 1309 1228 15.0576 β CH(Quin + Azomethyne) + β C—OH 1232 m — 1284 1205 117.6910 v C—OH + β CH(Quin + Azomethine) 1217 w 1220 w 1278 1199 32.0853 ν C═N(Py) + vC═C(Py) + β NH 1204 w 1204 vw 1267 1189 56.8637 β CH(Azo- methine + Quin) + β C—OH 1170 w 1172 sh 1253 1175 303.5474 βCH(Py) + βNH 1156 m 1156 m 1172 1099 380.9402 v N—N + βCH(Py + Quin) 1136 w 1133 w 1112 1043 10.8679 βCH(Py) 1122 sh — — 1105 w 1105 vw — 1090 w 1093 vw 1109 1040 19.7634 βCH(Py + Quin) 1072 w 1076 w 1092 1024 9.3109 βCH(Py) — 1063 vw 1079 1012 11.9293 βCH(Quin) + δ NNC 1044 vw 1044 vw 1068 1002 1.6514 βCH(Quin) 1007 vw — —  992 vw  992 w 1010 947 1.7715 Ring breath(Py)  981 vw — 1008 946 1.5921 γCH(Py)  950 vw  952 w 959 900 14.8080 γCH(Azomethine)  931 w  932 w 914 857 2.9777 Ring-deformation (Quin + Py)  895 w  897 vw —  881 vw — —  867 w  869 w 898 842 0.0665 γCH(Quin)  856 m  855 sh 880 826 11.8041 Ring-deformation (Quin + Py)  837 m  835 vw 892 837 2.8606 γCH(Py) —  812 vw 804 754 2.0048 γ C═C—C(Quin) + γ C═N—C(Quin)  792 vw  780 w 789 740 5.7350 β C═C—C(Quin + Azomethine)  766 s  768 vw 767 720 9.2858 γCH(Py)  720 m  720 m 734 689 18.7702 Ring- deformation(Quin)  696 w  696 vw 763 716 38.2602 γ CH(Quin)  677 s — 719 674 19.3631 β C═N—C(Py) + β C═C—C(Quin)  670 sh  668 w 698 655 2.0785 γ C═C—C(Quin) + β C═N—C(Py)  656 w  657 w 693 650 58.0279 Ring- deformation(Py)  644 sh — 681 639 1.5003 Ring- deformation(Py)  616 vw — 627 588 2.3638 Ring- deformation(Quin)  587 m — 604 567 90.9456 γ OH  575 w — —  547 sh  553 w 588 552 10.5255 β C═C—C(Quin)  532 sh  533 vw 559 524 1.6705 β C═C—C (Quin) + γ NH  522 w  523 vw 551 517 14.1393 γCH(Quin) + β C═C—C (Quin) + β C—OH  482 w  485 w 539/ 506/ 16.0281/ γ NH + 534 501 30.1104 γ C═C—C(Quin) Quin: quinoline ring; Py: pyridine ring; vs: very strong; s: strong; m: medium; w: weak; vw: very weak; br: broad; sh: shoulder; ip: in-phase; op: out-of-phase; v: stretching; β: in-plane bending; γ: out-of-plane bending. Carbonylic C═O stretching

The v C═O absorption is usually one of the most representative in an infrared spectrum and is also likely its most intense spectral feature. It appears in a wavenumber region relatively free of other vibrations (1800-1600 cm⁻¹). On the other hand, this mode gives only weak or very weak absorptions in Raman spectroscopy. In our study, as expected, v C═O vibration originates one of the strongest bands of the infrared spectrum, at 1656 cm⁻¹, which is in excellent agreement with the calculated value of 1658 cm⁻¹ (DFT calculations show a coupling between v C═O and β NH vibrations).

This mode was assigned at 1663 cm⁻¹ by Liu and Yang.

Azomethine C═N Stretching

The C═N stretchings of azomethine groups show absorptions close to that of carbonyl stretching. This fact can difficult an accurate assignment. For example, the C═N stretching bands of alkylated Schiff bases are usually found in the range 1674-1649 cm⁻¹, inside the common region of v C═O absorption. If conjugations of the C═N moiety with phenyl groups are present, the stretching frequency shifts to 1650-1600 cm⁻¹. In this work, two frequencies involving azomethine C═N vibrations were calculated (1569/1556 cm⁻¹), both of them coupled to v C═C of the quinoline ring and, to a lesser extent, to the phenol v C—OH. These values are in good agreement with the experimental frequencies observed in the infrared, at 1647 (vs) and 1604 (w) cm⁻¹, and Raman spectra, at 1646 (w) and 1603 (vs) cm⁻¹, respectively.

Liu and Yang, though, attributed this mode to a single band at 1613 cm⁻¹ in the IR spectrum, which was not observed in our study.

OH and NH stretching Vibrations

OH and NH groups are very characteristic and their stretching vibrations are observed, in many cases, around 3500-3300 cm⁻¹. This absorption, however, is highly influenced by chemical environment, mainly when OH or NH groups are involved in hydrogen bonding. This can occur within the same molecule (intramolecular H bonding) or with adjacent molecules (intermolecular H bonding). The presence of intramolecular H bonding causes a thinning of the band and makes its position unaffected by concentration changes. In the IR spectrum of INHHQ, we observed a sharp band of medium intensity located at 3396 cm⁻¹, assigned to v OH. A similar absorption, at 3418 cm⁻¹, was reported by Krishnakumar and Ramasamy in the infrared spectrum of 8-hydroxyquinoline (8-HQ). On the other hand, intermolecular hydrogen bonding usually leads to a broadening of the band, as can be seen in the case of the v NH absorption of INHHQ, which was attributed to the weak IR band at 3183 cm⁻¹. In a previous study on the isonicotinoyl hydrazone of 2-hydroxy-3-methoxybenzaldehyde, published by us, v NH vibration was observed as a weak band at 3157 cm⁻¹. Here, we found serious discrepancies concerning the assignments made by Liu and Yang, since these authors attributed an absorption of higher frequency (reported by them at 3576 cm⁻¹) to the NH stretching mode, whereas the lower frequency band at 3193 cm⁻¹ was credited to the OH stretching movement.

Phenol C—OH Vibrations

In this work, the C—OH stretching mode was assigned to the medium intensity infrared band at 1232 cm⁻¹. This vibration is Raman inactive and had its frequency calculated at 1205 cm⁻¹. A coupled mode involving this movement was also predicted at 1276 cm⁻¹ [experimental: 1299 (infrared) cm⁻¹ and 1304 (Raman) cm⁻¹]. Another important vibration concerning the phenol group is the in-plane bending, which typically appears in the region 1440-1260 cm⁻¹, attributed to the weak infrared band (medium in the Raman spectrum) at 1371 cm⁻¹. Coupled modes are observed in FTIR at 1465, 1280, 1252 and 1202 cm⁻¹. Theoretical and experimental frequencies show good agreement (Table 4).

Biological Activity Studies

This hydrazonic compound has the ability, as proved by 1D and 2D NMR experiments, of compete with the β-amyloid peptide (or α-synuclein), key-targets, respectively, in Alzheimer's and Parkinson's diseases, by physiological ions such as Zn²⁺ Cu²⁺ and Cu⁺, which can cause their precipitation or oligomerization and contribute to worsen the oxidative stress condition observed in the brains of patients suffering from these neurodegenerative disorders. INHHQ does not interact directly with the β-amyloid peptide and α-synuclein, nonetheless inhibits the interactions between these targets and metals through a mechanism that probably involves metal ion sequestering.

In silico pharmacological analyses (mandatory studies in the development of new therapeutic agents, whose aim is to predict pharmacokinetic properties of novel molecules with a potential pharmacological action) show that the Lipinski's parameters presented by INHHQ (Table 5), calculated via 1D-QSAR method, are in agreement to ideal values, suggesting that this compound constitutes an excellent candidate to new drug, with good oral absorption and cellular permeability. The model also indicates that INHHQ possesses structural features which can allow it crossing the blood-brain barrier (BBB), resulting in therapeutic actions inside the patients' brains.

TABLE 5 Lipinski parameters calculated for INHHQ, along with reference values Ref. Oral Ref. BBB Parameters INHHQ Biodisponibility Crossing HBD 2 ≤5 ≤3 HBA 6 ≤10 ≤7 MW 292 ≤500 ≤400 log P 2.34 −1 a 5 −1 a 5 log D (pH = 7.4) 2.27 −1 a 5 −1 a 5 log S −3.36 −4 a 2 −4 a 2 PSA 66.077 Å ≤140 Å ≤90 Å Rotatable bonds 4 ≤10 ≤10

Furthermore, both the INHHQ molecule itself and its potential metabolites were shown, in theoretical comparison with the toxic fragments of over 3000 comercially available drugs, completely non-toxic.

In fact, the intraperitoneal injection of up to 300 mg kg⁻¹ of the compound, using 10% DMSO/saline solution as vehicle of injection, in Wistar male rats (acute toxicity test) showed that INHHQ was apparently not toxic to the animals throughout the 72 h of the experiment: no animals died and there were no behavioral changes noted in the injected rats. Also, after the animals' sacrifice, there were no macroscopic abnormalities observed during its organs dissection. 

What is claimed is:
 1. A method for preparing medicaments for the treatment of neurodegenerative disorders, comprising using an anhydrous crystalline polymorph as an active drug in the medicament, the anhydrous crystalline polymorph comprising: a) the crystal structure of FIG. 1; b) crystal packing comprising at least one feature selected from the group consisting of: i) maintained by intermolecular H bonds involving the carbonyl oxygen O2 (acceptor) of a molecule and the N2-H group of the following molecule [moderate N2

O2^(i)=2,966 Å, symmetry codes: (i) −x+½, y−½, z], connecting the molecules of INHHQ in zigzag chains; or ii) molecules in each chain interconnected by cross-stacking π-π interactions involving the quinoline rings, being the calculated centroid-centroid distance equal to 3.8303(9) Å, and the adjacent chains interconnected by O1-H12

π interactions; or ii) as a result of the mentioned O1-H12

π interactions, zigzag columns run parallel to the crystallographic axisa; and c) the following characteristics: Melting point 246−249° C. Elemental analysis C, 66.3%; H, 4.1%; N, 19.4% Crystal system Orthorhombic a = 17.0761(4) Å Unit cell size  b = 8.25480(10) Å c = 19.3549(4) Å Z 8 ρ (calculated) 1.423 g cm⁻³


2. The method according to claim 1, wherein the anhydrous crystalline polymorph presents X-ray diffraction peaks (2θ ranging from 10.5 to 70.5°) as follows: Plane Multi- Interplanar 2 (h, k, l) plicity distance (Å) theta (°) H k l m d 1 0 0 2 8.34877 10.58783 0 1 0 2 7.07920 12.49361 0 1 1 4 5.94931 14.87873 1 0 −2 2 5.93863 14.90564 1 −1 −1 4 5.50433 16.08920 0 0 2 2 5.48852 16.13588 1 1 0 4 5.39942 16.40394 1 −1 −2 4 4.54974 19.49495 1 1 1 4 4.37713 20.27167 2 0 −2 2 4.37659 20.27421 0 1 2 4 4.33757 20.45853 2 0 0 2 4.17439 21.26742 2 −1 −1 4 3.87923 22.90666 1 0 2 2 3.87110 22.95541 2 −1 −2 4 3.72262 23.88427 2 1 0 4 3.59579 24.73979 0 2 0 2 3.53960 25.13890 1 −1 −3 4 3.53090 25.20189 1 1 2 4 3.39646 26.21681 0 2 1 4 3.36879 26.43604 1 −2 −1 4 3.28146 27.15290 1 2 0 4 3.25881 27.34522 2 −1 −3 4 3.25355 27.39035 0 1 3 4 3.25049 27.41659 3 0 −2 2 3.08971 28.87348 2 1 1 4 3.08859 28.88417 1 −2 −2 4 3.04049 29.35123 1 0 −4 2 3.03231 29.43221 1 2 1 4 2.98732 29.88574 0 2 2 4 2.97465 30.01599 2 0 −4 2 2.96931 30.07124 3 −1 −2 4 2.83175 31.56923 2 −2 −1 4 2.81367 31.77739 3 −1 −1 4 2.78865 32.07025 1 −1 −4 4 2.78737 32.08534 2 0 2 2 2.78466 32.11737 3 0 0 2 2.78292 32.13796 2 −2 −2 4 2.75217 32.50702 0 0 4 2 2.74426 32.60332 2 −1 −4 4 2.73820 32.67747 2 2 0 4 2.69971 33.15667 3 −1 −3 4 2.69759 33.18355 1 1 3 4 2.69411 33.22766 1 −2 −3 4 2.67192 33.51166 1 2 2 4 2.61222 34.30092 3 0 −4 2 2.61018 34.32857 2 1 2 4 2.59139 34.58541 3 1 0 4 2.58999 34.60470 0 1 4 4 2.55873 35.04100 2 −2 −3 4 2.54551 35.22898 0 2 3 4 2.54404 35.24991 2 2 1 4 2.46414 36.43237 3 −1 −4 4 2.44902 36.66531 0 3 0 2 2.35973 38.10499 3 −2 −2 4 2.32767 38.65065 1 0 4 2 2.32176 38.75283 4 0 −2 2 2.31874 38.80530 3 1 1 4 2.31772 38.82317 0 3 1 4 2.30703 39.01032 3 −2 −1 4 2.30355 39.07167 1 −2 −4 4 2.30283 39.08438 2 −1 −5 4 2.30097 39.11728 1 −3 −1 4 2.27839 39.52090 2 −2 −4 4 2.27487 39.58469 1 −1 −5 4 2.27151 39.64563 1 3 0 4 2.27077 39.65907 3 −2 −3 4 2.25142 40.01449 1 2 3 4 2.24939 40.05204 1 1 4 4 2.20614 40.87197 4 −1 −2 4 2.20355 40.92216 1 −3 −2 4 2.19295 41.12888 2 2 2 4 2.18856 41.21509 4 0 −4 2 2.18829 41.22042 3 2 0 4 2.18772 41.23171 4 −1 −3 4 2.18705 41.24487 2 1 3 4 2.18427 41.29979 1 3 1 4 2.17275 41.52890 0 2 4 4 2.16879 41.60831 3 −1 −5 4 2.16859 41.61228 0 3 2 4 2.16786 41.62688 4 −1 −1 4 2.13480 42.30230 3 0 2 2 2.13258 42.34844 2 −3 −1 4 2.10312 42.97079 3 −2 −4 4 2.10076 43.02150 0 1 5 4 2.09689 43.10495 4 −1 −4 4 2.09069 43.23919 4 0 0 2 2.08719 43.31522 2 −3 −2 4 2.07706 43.53728 2 3 0 4 2.05423 44.04625 3 1 2 4 2.04194 44.32549 1 −3 −3 4 2.04191 44.32613 2 0 −6 2 2.03690 44.44102 3 2 1 4 2.01612 44.92393 1 3 2 4 2.01489 44.95273 2 −2 −5 4 2.00506 45.18528 4 1 0 4 2.00199 45.25840 1 −2 −5 4 1.98548 45.65592 2 −3 −3 4 1.98380 45.69690 0 3 3 4 1.98310 45.71378 3 0 −6 2 1.97954 45.80066 1 0 −6 2 1.97817 45.83434 2 −1 −6 4 1.95748 46.34679 2 3 1 4 1.94455 46.67316 4 −1 −5 4 1.94222 46.73259 1 2 4 4 1.94138 46.75385 4 −2 −2 4 1.93962 46.79894 2 0 4 2 1.93555 46.90307 4 −2 −3 4 1.92834 47.08917 2 2 3 4 1.92643 47.13862 3 −2 −5 4 1.91564 47.42022 3 −1 −6 4 1.90641 47.66401 1 −1 −6 4 1.90518 47.69668 4 −2 −1 4 1.89223 48.04363 3 −3 −2 4 1.87535 48.50388 2 1 4 4 1.86703 48.73407 0 2 5 4 1.86568 48.77150 3 −3 −1 4 1.86266 48.85561 1 −3 −4 4 1.86228 48.86626 4 −2 −4 4 1.86131 48.89351 1 1 5 4 1.85776 48.99309 2 −3 −4 4 1.84740 49.28595 5 0 −2 2 1.83827 49.54725 4 1 1 4 1.83748 49.56990 3 −3 −3 4 1.83478 49.64793 1 3 3 4 1.83368 49.67962 4 0 −6 2 1.83168 49.73747 0 0 6 2 1.82951 49.80075 5 0 −4 2 1.82704 49.87241 3 2 2 4 1.82666 49.88361 2 3 2 4 1.80028 50.66584 3 3 0 4 1.79981 50.68000 4 2 0 4 1.79790 50.73774 5 −1 −3 4 1.79778 50.74113 3 1 3 4 1.79572 50.80350 0 3 4 4 1.78922 51.00132 5 −1 −2 4 1.77926 51.30747 4 −1 −6 4 1.77329 51.49295 0 1 6 4 1.77131 51.55465 0 4 0 2 1.76980 51.60187 5 −1 −4 4 1.76908 51.62454 2 −2 −6 4 1.76545 51.73845 4 −2 −5 4 1.75423 52.09423 3 −3 −4 4 1.75045 52.21513 0 4 1 4 1.74724 52.31836 1 −4 −1 4 1.73470 52.72556 1 4 0 4 1.73133 52.83614 3 −2 −6 4 1.72771 52.95534 1 −2 −6 4 1.72680 52.98560 5 −1 −1 4 1.71763 53.29042 4 0 2 2 1.71625 53.33669 3 3 1 4 1.70057 53.86813 5 −1 −5 4 1.69944 53.90672 2 2 4 4 1.69823 53.94829 1 −4 −2 4 1.69608 54.02211 2 −3 −5 4 1.69392 54.09682 1 2 5 4 1.69125 54.18921 2 −1 −7 4 1.69108 54.19489 1 4 1 4 1.68669 54.34788 0 4 2 4 1.68440 54.42786 1 −3 −5 4 1.68206 54.50961 3 −1 −7 4 1.68013 54.57753 4 2 1 4 1.67591 54.72649 5 0 0 2 1.66975 54.94516 4 1 2 4 1.66794 55.01012 1 3 4 4 1.65500 55.47716 4 −3 −2 4 1.65390 55.51703 2 −4 −1 4 1.65348 55.53225 4 −3 −3 4 1.64689 55.77391 2 3 3 4 1.64570 55.81771 5 −2 −3 4 1.64562 55.82063 5 0 −6 2 1.64485 55.84915 3 2 3 4 1.64404 55.87900 1 0 6 2 1.64248 55.93667 2 −4 −2 4 1.64073 56.00177 3 −3 −5 4 1.63896 56.06742 1 −1 −7 4 1.63562 56.19224 5 −2 −2 4 1.63138 56.35117 2 4 0 4 1.62941 56.42557 3 0 4 2 1.62858 56.45687 4 −2 −6 4 1.62677 56.52512 0 2 6 4 1.62525 56.58301 5 1 0 4 1.62516 56.58630 4 −3 −1 4 1.62423 56.62165 5 −2 −4 4 1.62352 56.64860 1 −4 −3 4 1.62324 56.65939 2 1 5 4 1.62060 56.76011 1 4 2 4 1.60956 57.18482 0 3 5 4 1.60734 57.27112 4 −1 −7 4 1.60639 57.30817 4 −3 −4 4 1.60455 57.38024 5 −1 −6 4 1.60217 57.47322 1 1 6 4 1.59998 57.55912 2 −4 −3 4 1.59358 57.81228 0 4 3 4 1.59322 57.82655 3 1 4 4 1.58712 58.06987 5 −2 −1 4 1.58349 58.21592 3 3 2 4 1.58219 58.26825 2 4 1 4 1.57302 58.64091 5 −2 −5 4 1.56920 58.79766 4 3 0 4 1.56339 59.03785 2 −2 −7 4 1.56261 59.07000 3 −2 −7 4 1.55396 59.43196 6 0 −4 2 1.54485 59.81773 4 2 2 4 1.54429 59.84161 2 −3 −6 4 1.54191 59.94356 3 −4 −2 4 1.53571 60.21080 4 −3 −5 4 1.53442 60.26657 0 1 7 4 1.53103 60.41363 3 −4 −1 4 1.52872 60.51473 1 −4 −4 4 1.52851 60.52394 3 0 −8 2 1.52498 60.67867 2 −4 −4 4 1.52025 60.88755 1 −2 −7 4 1.51854 60.96338 6 0 −2 2 1.51698 61.03271 3 −3 −6 4 1.51658 61.05053 5 1 1 4 1.51638 61.05931 2 0 −8 2 1.51616 61.06934 1 −3 −6 4 1.51596 61.07813 3 −4 −3 4 1.51319 61.20197 1 4 3 4 1.51257 61.22953 6 −1 −3 4 1.51024 61.33418 5 2 0 4 1.51016 61.33806 6 −1 −4 4 1.50933 61.37515 4 1 3 4 1.50872 61.40297 2 2 5 4 1.50649 61.50344 2 3 4 4 1.49652 61.95818 4 −2 −7 4 1.49506 62.02536 2 4 2 4 1.49366 62.09007 3 4 0 4 1.49339 62.10246 1 3 5 4 1.49174 62.17898 5 −2 −6 4 1.49166 62.18267 5 −1 −7 4 1.49157 62.18692 3 −1 −8 4 1.49078 62.22328 1 2 6 4 1.48989 62.26457 0 4 4 4 1.48733 62.38401 4 0 −8 2 1.48466 62.50882 6 −1 −2 4 1.48331 62.57221 2 −1 −8 4 1.48254 62.60831 4 3 1 4 1.48118 62.67218 6 −1 −5 4 1.48072 62.69367 3 2 4 4 1.47949 62.75186 3 −4 −4 4 1.46483 63.45251 1 0 −8 2 1.46062 63.65703 5 −3 −3 4 1.46015 63.67986 3 3 3 4 1.45904 63.73375 6 0 −6 2 1.45886 63.74255 2 0 6 2 1.45666 63.85023 4 −1 −8 4 1.45305 64.02798 5 −3 −2 4 1.45017 64.17003 4 −3 −6 4 1.44693 64.33096 0 3 6 4 1.44586 64.38455 5 −3 −4 4 1.44464 64.44527 3 4 1 4 1.43522 64.91979 0 2 7 4 1.43374 64.99512 6 −1 −1 4 1.43271 65.04772 5 0 2 2 1.43178 65.09530 2 −4 −5 4 1.43122 65.12385 1 −1 −8 4 1.43049 65.16140 6 −1 −6 4 1.42884 65.24586 2 1 6 4 1.42677 65.35220 1 −4 −5 4 1.42405 65.49281 5 2 1 4 1.42169 65.61510 6 −2 −3 4 1.41663 65.87928 5 −3 −1 4 1.41622 65.90063 6 −2 −4 4 1.41588 65.91867 0 5 0 2 1.41584 65.92052 4 2 3 4 1.41537 65.94542 3 1 5 4 1.41356 66.04055 1 4 4 4 1.40751 66.36081 4 −4 −2 4 1.40684 66.39668 5 −3 −5 4 1.40597 66.44291 5 0 −8 2 1.40574 66.45520 0 5 1 4 1.40421 66.53712 5 1 2 4 1.40336 66.58237 1 1 7 4 1.40288 66.60856 4 −4 −3 4 1.40251 66.62790 2 4 3 4 1.40178 66.66737 2 −3 −7 4 1.40123 66.69715 5 −2 −7 4 1.40118 66.69978 3 −2 −8 4 1.40053 66.73479 1 −5 −1 4 1.39767 66.88894 3 −4 −5 4 1.39761 66.89248 1 5 0 4 1.39591 66.98464 3 −3 −7 4 1.39498 67.03543 6 −2 −2 4 1.39432 67.07096 2 −2 −8 4 1.39368 67.10576 4 0 4 2 1.39233 67.17963 6 −2 −5 4 1.39218 67.18803 6 0 0 2 1.39146 67.22715 4 −4 −1 4 1.38844 67.39312 4 3 2 4 1.38797 67.41877 5 −1 −8 4 1.37882 67.92709 0 4 5 4 1.37785 67.98154 1 −5 −2 4 1.37724 68.01559 4 −4 −4 4 1.37608 68.08057 1 5 1 4 1.37219 68.30019 0 0 8 2 1.37213 68.30382 0 5 2 4 1.37096 68.37016 1 −3 −7 4 1.36918 68.47118 4 −2 −8 4 1.36910 68.47589 4 1 4 4 1.36616 68.64389 6 1 0 4 1.36534 68.69093 5 3 0 4 1.36303 68.82355 3 4 2 4 1.36190 68.88850 6 −1 −7 4 1.36065 68.96085 2 3 5 4 1.36033 68.97921 2 −5 −1 4 1.35414 69.33989 6 −2 −1 4 1.35205 69.46230 4 −3 −7 4 1.35190 69.47100 1 −2 −8 4 1.35018 69.57237 4 4 0 4 1.34986 69.59140 5 −3 −6 4 1.34938 69.61938 6 −2 −6 4 1.34879 69.65417 1 3 6 4 1.34807 69.69667 2 −5 −2 4 1.34710 69.75419 0 1 8 4 1.34706 69.75684 2 2 6 4 1.34705 69.75717 2 5 0 4 1.34082 70.12915 3 3 4 4 1.34035 70.15689 1 −5 −3 4 1.33737 70.33644 2 −4 −6 4 1.33596 70.42175 3 2 5 4 1.33592 70.42439


3. The method according to claim 1, wherein the anhydrous crystalline polymorph comprises the infrared spectrum as shown in FIG.
 5. 4. The method according to claim 1, wherein the anhydrous crystalline polymorph presents the next IR vibrational frequencies: C═O stretching at 1656 cm⁻¹; and azomethine C═N stretching at 1647 cm⁻¹ and 1604 cm⁻¹; and OH stretching at 3396 cm⁻¹; and NH stretching at 3183 cm⁻¹.
 5. The method according to claim 4, wherein the anhydrous crystalline polymorph comprises the infrared spectrum as shown in FIG.
 5. 6. The method according to claim 1, for use in preparing specific medication for the treatment of a disease selected from the group consisting of Alzheimer's disease, Parkinson's disease, and Huntington's disease. 